黄杞苷对H2O2诱导SH-SY5Y细胞氧化应激损伤的保护作用

目的 探讨黄杞苷对H₂O₂诱导SH-SY5Y细胞损伤的保护作用与分子机制。方法 黄杞苷(5、10、20、40、80和160 μmol·L^-1)预处理SH-SY5Y细胞12 h,MTT法检测黄杞苷对H₂O₂诱导损伤后细胞活力的影响;通过氮蓝四唑(NBT)法总超氧化物歧化酶(SOD)活力检测试剂盒和总谷胱甘肽过氧化物酶(NADPH)检测试剂盒测定黄杞苷对H₂O₂诱导损伤后SOD和谷胱甘肽过氧化物酶(GSH-Px)活力的影响;通过流式细胞术对SH-SY5Y细胞进行活性氧(ROS)水平的测定。黄杞苷(20 μmol·L^-1)预处理SH-SY5Y细胞12 h,H₂O₂孵育6、12和24 h或用黄杞苷(5、10和20 μmol·L^-1)预处理细胞12 h,H₂O₂孵育12 h,采用Western blot法分别检测Nrf2及HO-1表达情况。结果 与模型组比较,黄杞苷能够提高SH-SY5Y细胞的活力,显著提高SOD和GSH水平,减少ROS产生(P<0.05)。黄杞苷能够促进Nrf2蛋白由细胞质向细胞核内转移,黄杞苷(20 μmol·L^-1)预处理SH-SY5Y细胞12 h时转移达到顶峰。H₂O₂损伤模型下,黄杞苷浓度依赖性地促Nrf2的核内转移(P<0.05),并促进下游蛋白HO-1的表达(P<0.05)。结论 黄杞苷对H₂O₂诱导SH-SY5Y细胞损伤具有保护作用,其作用机制与抗氧化,促进Nrf2、HO-1的表达有关。

Protective effect of engeletin on H2O2 induced oxidative stress injury in SH-SY5Y cells

Objective To investigate the protective effect and molecular mechanism of engeletin on SH-SY5Y cells injured by H₂O₂.Methods SH-SY5Y cells were pretreated with engeletin (5, 10, 20, 40, 80 and 160 μmol·L^-1) for 12 h. MTT assay was used to detect the effect of engeletin on cell viability after H₂O₂ induced injury. The effects of engeletin on the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH PX) after H₂O₂ induced injury were determined by NBT method and NADPH method; the levels of reactive oxygen species (ROS) in SH-SY5Y cells were determined by flow cytometry. SH-SY5Y cells were pretreated with engeletin (20 μmol·L^-1) for 12 h and incubated with H₂O₂ for 6 h, 12 h and 24 h, or pretreated with engeletin (5, 10 and 20 μmol·L^-1) for 12 h and incubated with H₂O₂ for 12 h. The expressions of Nrf2 and HO-1 were detected by Western blot. Results Engeletin could protect SH-SY5Y cells against H₂O₂ induced injury. Compared with the model group, engeletin could increase the cell viability, significantly increase the levels of SOD and GSH, and reduce the production of ROS (P<0.05). Engeletin promoted the transfer of Nrf2 protein from cytoplasm to nucleus, and reached the peak when SH-SY5Y cells were pretreated with engeletin (20 μmol·L^-1) for 12 h. In the model group of H₂O₂ induced injury, engeletin promoted the nuclear transfer of Nrf2 in a concentration dependent manner (P<0.05), and promoted the expression of HO-1, a downstream protein of Nrf2 (P<0.05).Conclusion Engeletin protected SH-SY5Y cells from H₂O₂ induced injury, and its mechanism was related to antioxidation and promoting the expression of Nrf2 and HO-1.